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Article ; Online: A gas phase fractionation acquisition scheme integrating ion mobility for rapid diaPASEF library generation.

Penny, Jack / Arefian, Mohammad / Schroeder, Gunnar N / Bengoechea, José A / Collins, Ben C

Proteomics

2023  Volume 23, Issue 7-8, Page(s) e2200038

Abstract: Data independent acquisition (DIA/SWATH) MS is a primary strategy in quantitative proteomics. diaPASEF is a recent adaptation using trapped ion mobility spectrometry (TIMS) to improve selectivity/sensitivity. Complex DIA spectra are typically analyzed ... ...

Abstract Data independent acquisition (DIA/SWATH) MS is a primary strategy in quantitative proteomics. diaPASEF is a recent adaptation using trapped ion mobility spectrometry (TIMS) to improve selectivity/sensitivity. Complex DIA spectra are typically analyzed with reference to spectral libraries. The best-established method for generating libraries uses offline fractionation to increase depth of coverage. More recently strategies for spectral library generation based on gas phase fractionation (GPF), where a representative sample is injected serially using narrow DIA windows that cover different mass ranges of the complete precursor space, have been introduced that performed comparably to deep offline fractionation-based libraries. We investigated whether an analogous GPF-based approach that accounts for the ion mobility (IM) dimension is useful for the analysis of diaPASEF data. We developed a rapid library generation approach using an IM-GPF acquisition scheme in the m/z versus 1/K0 space requiring seven injections of a representative sample and compared this with libraries generated by direct deconvolution-based analysis of diaPASEF data or by deep offline fractionation. We found that library generation by IM-GPF outperformed direct library generation from diaPASEF and had performance approaching that of the deep library. This establishes the IM-GPF scheme as a pragmatic approach to rapid library generation for analysis of diaPASEF data.
MeSH term(s) Proteomics/methods ; Peptide Library ; Chemical Fractionation/methods ; Proteome/analysis
Chemical Substances Peptide Library ; Proteome
Language English
Publishing date 2023-03-10
Publishing country Germany
Document type Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID 2032093-0
ISSN 1615-9861 ; 1615-9853
ISSN (online) 1615-9861
ISSN 1615-9853
DOI 10.1002/pmic.202200038
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Zs.A 5386: Show issues Location:
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Zs.A 1868: Show issues Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
Jg. 1995 - 2021: Lesesall (1.OG)
ab Jg. 2022: Lesesaal (EG)
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